Correlative microscopy and electron tomography of GFP through photooxidation.

Grabenbauer, Markus (author)

Geerts, Willie J C (author)

Fernandez-Rodriguez, Julia, 1965 (author): Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry

Hoenger, Andreas (author)

Koster, Abraham J (author)

Nilsson, Tommy, 1960 (author): Gothenburg University,Göteborgs universitet,Institutionen för medicinsk och fysiologisk kemi,Institute of Medical Biochemistry

(creator_code:org_t)

2005-10-21
2005
English.
In: Nature methods. - : Springer Science and Business Media LLC. - 1548-7091 .- 1548-7105. ; 2:11, s. 857-62

Related links:: https://gup.ub.gu.se...; show more...; https://doi.org/10.1...; show less...

Abstract Subject headings

We have developed a simple correlative photooxidation method that allows for the direct ultrastructural visualization of the green fluorescent protein (GFP) upon illumination. The method, termed GRAB for GFP recognition after bleaching, uses oxygen radicals generated during the GFP bleaching process to photooxidize 3,3'-diaminobenzidine (DAB) into an electron-dense precipitate that can be visualized by routine electron microscopy and electron tomography. The amount of DAB product produced by the GRAB method appears to be linear with the initial fluorescence, and the resulting images are of sufficient quality to reveal detailed spatial information. This is exemplified by the observed intra-Golgi stack and intracisternal distribution of a human Golgi resident glycosylation enzyme, N-acetylgalactosaminyltransferase-2 fused either to enhanced GFP or CFP.

By the author/editor: Grabenbauer, Mar ...; Geerts, Willie J ...; Fernandez-Rodrig ...; Hoenger, Andreas; Koster, Abraham ...; Nilsson, Tommy, ...

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